Human bocavirus 1(HBoV1;family:Parvoviridae)causes a wide spectrum of respiratory diseases in children and gastroenteritis in adults.A lack of sensitive cell lines and efficient animal models hinders research on HBoV,...Human bocavirus 1(HBoV1;family:Parvoviridae)causes a wide spectrum of respiratory diseases in children and gastroenteritis in adults.A lack of sensitive cell lines and efficient animal models hinders research on HBoV,including the development of anti-HBoV drugs or vaccines.Although the construction of a wild-type HBoV1 infectious clone has been reported,generating HBoV1 infectious clone carrying foreign reporter genes with suitable insertion sites in its genome while retaining replicative ability remains challenging.Here,HBoV1 infectious clones harboring the 11-amino-acid HiBiT tag at five distinct insertion sites were constructed and evaluated.Only the recombinant HBoV1 carrying the HiBiT tag in the N-terminus of the NS1 protein(HBoV1-HiBiTNS1)displayed comparable characteristics to wild-type HBoV1 as determined via the analysis of viral DNA copy number,NanoLuc activity,viral protein expression,and the formation of replication intermediates.Notably,the replication kinetics of HBoV1-HiBiTNS1 could be examined by monitoring NanoLuc activity,which was noted to be correlated with the viral DNA level.Additionally,we successfully applied HiBiT-tagged HBoV1 for the evaluation of antiviral drug activity and identified ivermectin(EC50=2.27μM)as a potent anti-HBoV1 replication drug.Overall,our study demonstrated that the HBoV1-HiBiTNS1 reporter can serve as a convenient platform for screening candidate drugs targeting HBoV1 replication and may also be useful for investigating the life cycle of the virus.展开更多
目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒...目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒PCR阳性率18.68%,其中flu A+flu B 10.44%,HRV 2.75%,ADV、HBo V和PV1型均为0.82%;RSVB 0.55%,PV2、PV4均为0.27%,hm PV未检出。比较一个流行年不同监测点流感检出率无差异。2012-2016年聚集性流感样病例疫情11起,集中在冬春季,流感占90.91%,无暴发疫情。结论徐汇地区呼吸道病毒多样化,流感病毒最常见,且呈冬春季和夏季双峰流行,B型、季H3型、新H1H1型交替,夏季高峰多以季H3型为优势毒株。流感样病例就诊百分比与流感病毒检出率呈正相关。学校、托幼机构为呼吸道病毒感染聚集疫情重点防控单位。应长期监测主要呼吸道病毒活动水平。展开更多
Background: Acute respiratory infection (ARI) is a leading cause of morbidity and mortality in children under five years of age in developing countries with viruses contributing significantly to this problem. The rece...Background: Acute respiratory infection (ARI) is a leading cause of morbidity and mortality in children under five years of age in developing countries with viruses contributing significantly to this problem. The recently identified parvovirus, Human Bocavirus (HBoV), has also been associated with ARI. Objective: To determine the frequency of HBoV in patients with ARI. Materials and Methods: Samples from 125 consenting patients with influenza like illness signs and symptoms were collected. DNA was extracted from these samples using the QIAamp DNA blood mini kit (Qiagen, Germany). Conventional PCR was carried out and the amplicons were examined in 2% agarose gels stained with ethidium bromide. This was followed by sequencing of the HBoV positive samples. Results: Twenty one (16.8%) patients were found to have HBoV infection. Males (n = 61.9%) were mainly infected with HBoV. Local HBoV strains had 98.9% - 100% similarities and were found to cluster together with other strains obtained elsewhere. Conclusion: These findings suggest that HBoV plays a role in respiratory tract infections in children in Kenya just like it has been found elsewhere. It also sheds light on multiple infections associated with HBoV infections in Kenya.展开更多
基金supported by the Guangzhou National Laboratory(SRPG22-002 to X.W.Chen)the Pearl River Talent Recruitment Program(2019CX01Y422 to X.W.Chen)+1 种基金the Natural Science Foundation of Guangdong Province(2025A1515011018 to J.L.Tang)the Basic and Applied Basic Research Projects of Guangzhou Basic Research Program(2025A04J5492 to J.L.Tang,2023A04J0161 to Q.Yang).
文摘Human bocavirus 1(HBoV1;family:Parvoviridae)causes a wide spectrum of respiratory diseases in children and gastroenteritis in adults.A lack of sensitive cell lines and efficient animal models hinders research on HBoV,including the development of anti-HBoV drugs or vaccines.Although the construction of a wild-type HBoV1 infectious clone has been reported,generating HBoV1 infectious clone carrying foreign reporter genes with suitable insertion sites in its genome while retaining replicative ability remains challenging.Here,HBoV1 infectious clones harboring the 11-amino-acid HiBiT tag at five distinct insertion sites were constructed and evaluated.Only the recombinant HBoV1 carrying the HiBiT tag in the N-terminus of the NS1 protein(HBoV1-HiBiTNS1)displayed comparable characteristics to wild-type HBoV1 as determined via the analysis of viral DNA copy number,NanoLuc activity,viral protein expression,and the formation of replication intermediates.Notably,the replication kinetics of HBoV1-HiBiTNS1 could be examined by monitoring NanoLuc activity,which was noted to be correlated with the viral DNA level.Additionally,we successfully applied HiBiT-tagged HBoV1 for the evaluation of antiviral drug activity and identified ivermectin(EC50=2.27μM)as a potent anti-HBoV1 replication drug.Overall,our study demonstrated that the HBoV1-HiBiTNS1 reporter can serve as a convenient platform for screening candidate drugs targeting HBoV1 replication and may also be useful for investigating the life cycle of the virus.
文摘目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒PCR阳性率18.68%,其中flu A+flu B 10.44%,HRV 2.75%,ADV、HBo V和PV1型均为0.82%;RSVB 0.55%,PV2、PV4均为0.27%,hm PV未检出。比较一个流行年不同监测点流感检出率无差异。2012-2016年聚集性流感样病例疫情11起,集中在冬春季,流感占90.91%,无暴发疫情。结论徐汇地区呼吸道病毒多样化,流感病毒最常见,且呈冬春季和夏季双峰流行,B型、季H3型、新H1H1型交替,夏季高峰多以季H3型为优势毒株。流感样病例就诊百分比与流感病毒检出率呈正相关。学校、托幼机构为呼吸道病毒感染聚集疫情重点防控单位。应长期监测主要呼吸道病毒活动水平。
文摘Background: Acute respiratory infection (ARI) is a leading cause of morbidity and mortality in children under five years of age in developing countries with viruses contributing significantly to this problem. The recently identified parvovirus, Human Bocavirus (HBoV), has also been associated with ARI. Objective: To determine the frequency of HBoV in patients with ARI. Materials and Methods: Samples from 125 consenting patients with influenza like illness signs and symptoms were collected. DNA was extracted from these samples using the QIAamp DNA blood mini kit (Qiagen, Germany). Conventional PCR was carried out and the amplicons were examined in 2% agarose gels stained with ethidium bromide. This was followed by sequencing of the HBoV positive samples. Results: Twenty one (16.8%) patients were found to have HBoV infection. Males (n = 61.9%) were mainly infected with HBoV. Local HBoV strains had 98.9% - 100% similarities and were found to cluster together with other strains obtained elsewhere. Conclusion: These findings suggest that HBoV plays a role in respiratory tract infections in children in Kenya just like it has been found elsewhere. It also sheds light on multiple infections associated with HBoV infections in Kenya.
